The objective of this proposal is to complete the development, engineering, and validation of the infectious disease diagnositics platform known as TIGER (Triangulation Identification for Genetic Evaluation of Risk), and to deploy a working version at the CDC. TIGER was created in a DARPA-sponsored program to develop technology to detect a broad range of biological weapons agents in samples collected from the environment. TIGER employs an electrospray ionization/mass spectrometyr-based detector coupled with high-throughput PCR and uses a maximum-likelihood data processor similar to those used for radar and satellite imaging applications. In tests involved hundreds of environmental samples, TIGER has successfully identified, without false positives, key biological weapons agents (spiked) such as Bacillus anthracis, Yersinia pestis, and Francisella tularensis. TIGER has also detected and strain-typed other medically important bacteria and viruses including Streptococcus pyogenes and Adenovirus in clinical samples. While TIGER has achieved critical proof of principle, the instrumentation needs further development and validation for NIAID/CDC-priority pathogens such as the Poxviridae, Arenaviridae, Bunyaviridae, Flaviviridae, Togaviridae (especially VEE), and Filoviridea families and the alpha, beta, gamma-proteobacteria. To enable deployment at the CDC, the mass spectrometry detector must be developed on an inexpensive and small sample clean-up, and PCR robotics must be consolidated on a dedicated instrument. Upon completion of the hardware engineering, thorough testing and validation of all biochemical reagents will be required. We believe that the most efficient route to broad utilization of TIGER technology is to develop the hardware, software, and biochemical reagents, then transition the technology to the CDC, who we will work with in a collaborative mode to develop important applications. Upon completion of these goals, the CDC will be capable of analyzing >100 of samples per day to detect and strain-type bacteria and viruses and perform high throughput multi-locus sequence typing (MLST) on the TIGER platform.